Mob-5/hmob-5 as a cancer diagnostic marker

Abstract

The invention provides isolated nucleic acids comprising the nucleic acids set forth in the Sequence Listing as SEQ ID NO: 1(corresponding to a cDNA encoding a rat Mob-5 protein), SEQ ID NO: 3(corresponding to a cDNA encoding the cancer specific human Mob-5 protein homolog with an internal deletion of 53 amino acid residues, referred to as cMob-5.), SEQ ID NO: 5(corresponding to a cDNA encoding a human Mob-5 protein), SEQ ID NO: 7(corresponding to a nucleic acid encoding a rat Mob-5-AP fusion protein and SEQ ID NO: 9(corresponding to a nucleic acid encoding a human Mob-5-AP fusion protein). The invention also provides purified polypeptides having the sequences set forth in the Sequence Listing as any of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10. This invention also relates to a method of detecting the presence of cancer in a patient comprising: a) contacting a sample from the patient with an antibody to Mob-5; and b) detecting the binding of the antibody with an antigen in the sample, wherein binding of antigen to the antibody indicates the presence of Mob-5 antigen in the sample and wherein Mob-5 antigen in the sample indicates the presence of cancer in the patient, thereby detecting the presence of cancer in the patient.

What is claimed is:  
     
         1 . An isolated nucleic acid comprising the nucleic acid set forth in the Sequence Listing as SEQ ID NO: 1.  
     
     
         2 . The isolated nucleic acid of  claim 1  in a vector suitable for expressing the nucleic acid.  
     
     
         3 . The vector of  claim 2  in a host suitable for expressing the nucleic acid.  
     
     
         4 . A purified polypeptide encoded by the nucleic acid of  claim 1 .  
     
     
         5 . An isolated nucleic acid encoding the polypeptide of  claim 4 .  
     
     
         6 . The polypeptide of  claim 4 , having the sequence set forth in the Sequence Listing as SEQ ID NO: 2.  
     
     
         7 . A purified antibody which specifically binds to the polypeptide of  claim 6 .  
     
     
         8 . An isolated nucleic acid comprising the nucleic acid set forth in the Sequence Listing as SEQ ID NO: 3.  
     
     
         9 . The isolated nucleic acid of  claim 8  in a vector suitable for expressing the nucleic acid .  
     
     
         10 . The vector of  claim 9  in a host suitable for expressing the nucleic acid.  
     
     
         11 . A purified polypeptide encoded by the nucleic acid of  claim 8 .  
     
     
         12 . An isolated nucleic acid encoding the polypeptide of  claim 11 .  
     
     
         13 . The polypeptide of  claim 12 , having the sequence set forth in the Sequence Listing as SEQ ID NO: 4.  
     
     
         14 . A purified antibody which specifically binds to the polypeptide of  claim 13 .  
     
     
         15 . An isolated nucleic acid comprising the nucleic acid set forth in the Sequence Listing as SEQ ID NO: 5.  
     
     
         16 . The isolated nucleic acid of  claim 15  in a vector suitable for expressing the nucleic acid.  
     
     
         17 . The vector of  claim 16  in a host suitable for expressing the nucleic acid.  
     
     
         18 . A purified polypeptide encoded by the nucleic acid of  claim 15 .  
     
     
         19 . An isolated nucleic acid encoding the polypeptide of  claim 18 .  
     
     
         20 . The polypeptide of  claim 18 , having the sequence set forth in the Sequence Listing as SEQ ID NO: 6.  
     
     
         21 . A purified antibody which specifically binds to the polypeptide of  claim 20 .  
     
     
         22 . The isolated nucleic acid of  claim 1  comprising the nucleic acid set forth in the Sequence Listing as SEQ ID NO: 7.  
     
     
         23 . The isolated nucleic acid of  claim 22  in a vector suitable for expressing the nucleic acid.  
     
     
         24 . The vector of  claim 23  in a host suitable for expressing the nucleic acid.  
     
     
         25 . A purified polypeptide encoded by the nucleic acid of  claim 22 .  
     
     
         26 . An isolated nucleic acid encoding the polypeptide of  claim 25 .  
     
     
         27 . The polypeptide of  claim 26 , having the sequence set forth in the Sequence Listing as SEQ ID NO: 8.  
     
     
         28 . A purified antibody which specifically binds to the polypeptide of  claim 27 .  
     
     
         29 . The isolated nucleic acid of  claim 15  comprising the nucleic acid set forth in the Sequence Listing as SEQ ID NO: 9.  
     
     
         30 . The isolated nucleic acid of  claim 29  in a vector suitable for expressing the nucleic acid.  
     
     
         31 . The vector of  claim 30  in a host suitable for expressing the nucleic acid.  
     
     
         32 . A purified polypeptide encoded by the nucleic acid of  claim 29 .  
     
     
         33 . An isolated nucleic acid encoding the polypeptide of  claim 32 .  
     
     
         34 . The polypeptide of  claim 32 , having the sequence set forth in the Sequence Listing as SEQ ID NO: 10.  
     
     
         35 . A purified antibody which specifically binds to the polypeptide of  claim 34 .  
     
     
         36 . A method of detecting the presence of cancer in a patient comprising: 
 a. contacting a sample from the patient with an antibody to Mob-5; and    b. detecting the binding of the antibody with an antigen in the sample, wherein binding of antigen to the antibody indicates the presence of Mob-5 antigen in the sample and wherein Mob-5 antigen in the sample indicates the presence of cancer in the patient, thereby detecting the presence of cancer in the patient.    
     
     
         37 . The method of  claim 36  wherein the cancer is a ras-induced cancer.  
     
     
         38 . The method of  claim 36 , wherein the cancer is a colorectal cancer.  
     
     
         39 . A method of detecting the presence of cancer in a patient comprising: 
 a. contacting a sample from the patient with Mob-5 antigen; and    b. detecting the binding of the antigen with an antibody in the sample, wherein binding of antibody to the antigen indicates the presence of Mob-5 antibody in the sample and wherein Mob-5 antibody in the sample indicates the presence of cancer in the patient, thereby detecting the presence of cancer in the patient.    
     
     
         40 . The method of  claim 39  wherein the cancer is a ras-induced cancer.  
     
     
         41 . The method of  claim 39 , wherein the cancer is a colorectal cancer.  
     
     
         42 . A method of determining the effectiveness of an anti-cancer therapy comprising: 
 a. obtaining a sample from a patient undergoing anti-cancer therapy, and b. monitoring the sample for expression of a mob-5 gene, whereby inhibition of the expression of the mob-5 gene indicates the anti-cancer therapy is effective, thereby determining the effectiveness of an anti-cancer therapy.    
     
     
         43 . A method of determining the effectiveness of an anti-cancer therapy comprising: 
 a. obtaining a sample from a patient undergoing anti-cancer therapy, and    b. monitoring the sample for active mob-5 gene product, whereby inhibition of the activity of the mob-5 gene product indicates the anti-cancer therapy is effective, thereby determining the effectiveness of an anti-cancer therapy.    
     
     
         44 . A method of screening agents for anti-cancer activity comprising; 
 a. administering the agent to a cancer cell;    b. monitoring the expression of a mob-5 gene in the cell, whereby an inhibition of the expression of the mob-5 gene indicates the agent has anti-cancer activity, thereby screening the agent for anti-cancer activity.    
     
     
         45 . A method of screening agents for anti-cancer activity comprising; 
 a. administering the agent to a cancer cell;    b. monitoring the activity of a Mob-5 gene product in the cell, whereby an inhibition of the activity of the Mob-5 gene product indicates the agent has anti-cancer activity, thereby screening the agent for anti-cancer activity.    
     
     
         46 . A method of inhibiting a cellular transformation phenotype induced by an oncogene whose product functions upstream of a mob-5 gene product, comprising inhibiting formation of an active mob-5 gene product in a cell containing the upstream oncogene so as to inhibit the expression of the cellular transformation phenotype of the upstream oncogene.  
     
     
         47 . The method of  claim 46 , wherein the cellular transformation phenotype is inhibited by a nucleic acid antisense to the mob-5 gene.  
     
     
         48 . The method of  claim 46 , wherein the cellular transformation phenotype is induced by a ras oncogene product.  
     
     
         49 . A method of inhibiting a cellular transformation phenotype induced by an oncogene whose product functions upstream of a mob-5 gene product, comprising inhibiting an active mob-5 gene product in a cell containing the upstream oncogene so as to inhibit the expression of the cellular transformation phenotype of the upstream oncogene.  
     
     
         50 . The method of  claim 49 , wherein the mob-5 gene product is inhibited by an antibody to the mob-5 gene product.  
     
     
         51 . The method of  claim 49 , wherein the cellular transformation phenotype is induced by a ras oncogene product.  
     
     
         52 . A method for inhibiting a mob-5 induced cellular transformation phenotype comprising inhibiting the formation of active Mob-5 gene product in a cell so as to inhibit the expression of the mob-5 induced cellular transformation phenotype.  
     
     
         53 . The method of  claim 52 , wherein the inhibition of the formation of an active Mob-5 protein is by a nucleic acid antisense to mob-5.  
     
     
         54 . A method for inhibiting a mob-5 induced cellular transformation phenotype comprising inhibiting an active Mob-5 gene product in a cell so as to inhibit the expression of the mob-5 induced cellular transformation phenotype.  
     
     
         55 . The method of  claim 54 , wherein the active Mob-5 gene product is inhibited by an antibody to Mob-5.  
     
     
         56 . A method of determining the effectiveness of an anti-cancer therapy comprising: 
 a. obtaining a sample from a patient undergoing anti-cancer therapy, and    b. monitoring the sample for the receptor for the mob-5 gene product, whereby a decreased amount of the receptor for the mob-5 gene product indicates the anti-cancer therapy is effective, thereby determining the effectiveness of an anti-cancer therapy.    
     
     
         57 . A method of determining the effectiveness of an anti-cancer therapy comprising: 
 a. obtaining a sample from a patient undergoing anti-cancer therapy, and    b. monitoring the sample for expression of a gene encoding a Mob-5 receptor, whereby inhibition of the expression of the gene encoding the Mob-5 receptor indicates the anti-cancer therapy is effective, thereby determining the effectiveness of an anti-cancer therapy.    
     
     
         58 . A method of screening agents for anti-cancer activity comprising; 
 a. administering the agent to a cancer cell;    b. monitoring the expression of a gene encoding a Mob-5 receptor in the cell, whereby an inhibition of the expression of the gene encoding a Mob-5 receptor indicates the agent has anti-cancer activity, thereby screening the agent for anti-cancer activity.    
     
     
         59 . A method of screening agents for anti-cancer activity comprising; 
 a. administering the agent to a cancer cell;    b. monitoring the activity of a Mob-5 receptor in the cell, whereby an inhibition of the activity of the Mob-5 receptor indicates the agent has anti-cancer activity, thereby screening the agent for anti-cancer activity.    
     
     
         60 . A method of inhibiting a cellular transformation phenotype induced by an oncogene whose product functions upstream of a mob-5 gene product, comprising inhibiting formation of an active Mob-5 receptor in a cell containing the upstream oncogene so as to inhibit the expression of the cellular transformation phenotype of the upstream oncogene.  
     
     
         61 . The method of  claim 60 , wherein the cellular transformation phenotype is inhibited by a nucleic acid antisense to a Mob-5 receptor gene.  
     
     
         62 . The method of  claim 60 , wherein the cellular transformation phenotype is induced by a ras oncogene product.  
     
     
         63 . A method of inhibiting a cellular transformation phenotype induced by an oncogene whose product functions upstream of a mob-5 gene product, comprising inhibiting an active Mob-5 receptor in a cell containing the upstream oncogene so as to inhibit the expression of the cellular transformation phenotype of the upstream oncogene.  
     
     
         64 . The method of  claim 63 , wherein the Mob-5 receptor is inhibited by an antibody to the Mob-5 receptor.  
     
     
         65 . The method of  claim 63 , wherein the cellular transformation phenotype is induced by a ras oncogene product.  
     
     
         66 . A method for inhibiting a mob-5 induced cellular transformation phenotype comprising inhibiting the formation of active Mob-5 receptor in a cell so as to inhibit the expression of the mob-5 induced cellular transformation phenotype.  
     
     
         67 . The method of  claim 66 , wherein the inhibition of the formation of an active Mob-5 receptor is by a nucleic acid antisense to the Mob-5 receptor gene.  
     
     
         68 . A method for inhibiting a mob-5 induced cellular transformation phenotype comprising inhibiting an active Mob-5 receptor in a cell so as to inhibit the expression of the mob-5 induced cellular transformation phenotype.  
     
     
         69 . The method of  claim 68 , wherein the active Mob-5 receptor is inhibited by an antibody to the Mob-5 receptor.  
     
     
         70 . The method of  claim 68 , wherein the active Mob-5 receptor is inhibited by administering to the cell an altered Mob-5 which binds to the Mob-5 receptor, whereby binding of the altered Mob-5 to the receptor inhibits the binding of Mob-5 to the receptor, thereby inhibiting the expression of the mob-5 induced cellular transformation phenotype.  
     
     
         71 . A method of detecting the presence of cancer in a patient comprising: 
 a. contacting a sample from the patient with an antibody to a Mob-5 receptor;    b. detecting the binding of the antibody with an antigen in the sample, wherein binding of antigen to the antibody indicates the presence of Mob-5 receptor antigen in the sample and wherein Mob-5 receptor antigen in the sample indicates the presence of cancer in the patient, thereby detecting the presence of cancer in the patient.    
     
     
         72 . The method of  claim 71 , wherein the cancer is a ras-induced cancer.  
     
     
         73 . The method of  claim 71 , wherein the cancer is a colorectal, cancer.  
     
     
         74 . A method of detecting the presence of cancer in a patient comprising: 
 a. contacting a sample from the patient with a Mob-5 receptor antigen;    b. detecting the binding of the antigen with an antibody in the sample, wherein binding of antigen to the antibody indicates the presence of Mob-5 receptor antibody in the sample and wherein Mob-5 receptor antibody in the sample indicates the presence of cancer in the patient, thereby detecting the presence of cancer in the patient.    
     
     
         75 . The method of  claim 74 , wherein the cancer is a ras-induced cancer.  
     
     
         76 . The method of  claim 74 , wherein the cancer is a colorectal, cancer.  
     
     
         77 . A method of detecting the Mob-5 receptor comprising, contacting a cell with a Mob5-AP fusion protein, detecting the binding of the Mob5-AP fusion protein to the cell, wherein binding of the Mob5-AP to the cell indicates the cell is a Mob-5 receptor expressing cell, thus detecting the Mob-5 receptor.  
     
     
         78 . A method of detecting the presence of cancer in a patient comprising: 
 a. contacting a sample from the patient with a mixture of biotinylated monoclonal antibody to Mob-5 and Streptavidin-horseradish peroxidase (HRP);    b. adding Gel Blue A Sepharose beads to the sample of a);    c. spinning down the beads;    d. removing supernatant;    e. adding HRP substrate    f. detecting the presence of color, wherein the presence of color indicates the presence of Mob-5 antigen in the sample and wherein Mob-5 antigen in the sample indicates the presence of cancer in the patient, thereby detecting the presence of cancer in the patient.    
     
     
         79 . The method of  claim 78 , wherein the antibody to Mob-5 is the monoclonal antibody produced from hybridoma clone L-14.  
     
     
         80 . The antibody of  claim 21 , wherein the antibody is a monoclonal antibody produced from hybridoma clone L-14.